Ruminant feed composition containing an organic acid amide



United States Patent RUMINANT FEED COMPOSITION CONTAINING AN ORGANIC ACID AMIDE Irvin J. Belasco and Kenneth E. Walker, Wilmington,

Del., assignors to E. I. du Pont de Nemours and Company, Wilmington, Del., a corporation of Delaware No Drawing. Application November 3, 1951, Serial No. 254,788

Claims. (Cl. 9 9-2) This invention relates to a new source of nitrogen to supplement the protein feed ofRumin'antia, a division of ungulate mammals, with a quadripartite stomach and more especially is directed to providing amides of organic acids as a source of usable nitrogen to supplement the protein feeds for such animals.

Burroughs et al., of the Ohio Agricultural Experimental Station, have called attention to the importance of nutrients needed by rumen microorganisms in the digestion of roughages and particularly the cellulose fraction of their normal feed. (Jour. Animal Science, 9, 523-530, 1950). From the fact that .the feeding of cattle and other ruminants often includes grains and protein-rich feeds in combinationwith roughage, Burroughs has postulated that these grains and protein-rich feeds, when fed with roughage, might supply nutritional factors needed by rumen microorganisms in the, digestion of the roughage fraction of the ration, (Feedstuffs, p. 24, 12-9-50). These investigators present considerable evidence supporting the proposition that grains and other protein-rich feeds contribute to the nutritionof rumen microorganisms in their digestion of cellulose.

Further Work has established that urea'is a source of usable nitrogen which may-replace a portion, at least, of the protein feed for such animals. By usable nitrogen is meant the nitrogen of a compound that is a food for rumen microflora. An improved use of compounds derived from urea is described in the copending application of M. F. Gribbins, S. N. 196,707, filed November 20, 1950. In the use of urea as a source of usable nitrogen, care must be exercised to control carefully the amount of urea available to the animal. It has been found that urea and its derivatives canbe used to supply to and preferably about /3 of the protein requirements. Inasmuch as, however, large quantities of urea per se under certain conditions are broken down by microorganisms and.amidasespresentinthe rumen of certain animals, giving free ammonia, care must be exercised to prevent excess formation of the ammonia in toxic concentrations. (J. S. Dinning and H. M. Briggs, Am. J. Physiol. 153, 41-46 (1948)).

An object of the present invention is to provide a new source of usable nitrogen to supplement the protein feed for the microflora of. Ruminantia. .Yet anotherobject is to provide from'amides of organic acids, as sources of nitrogen, supplemental. feeds. in .which the usable synthetic nitrogen ration of the animal maybe increased and the potential hazards resulting from ammonia evolution minimized. Still another object is to provide a supplemental feed composition for cattle containing, as an essential ingredient thereof, certain amides of the organic acids. A further object is to provide organic acid amides readily available for utilization by rumen microorganisrris. Other objects and advantages of the invention:

will hereinafter 'appe'ar. v v.

In accord with the invention it has beenfound that amides of organic acidsgenerally can be used asa source of nitrogen to supplementprotein feed of Ruminantia with a minimum requirement for restricting the ingestion of the amide by the animal. While the said amides break down more slowly to provide nutrients for the rumen microorganisms in the digestion of roughages, and more particularly cellulose in its various forms, the breakdown rate to ammonia is considerably less than that from other synthetic nitrogen compounds such as urea, for example.

Work with cattle and sheep, in order to determine the value of feeds generally, has been conducted until recently directly by feeding the animal. Within the past few years an in vitro rumen and technique for its use have been developed whereby the changes taking place in feeds, brought about by microorganisms, can be measured more readily and accurately. This technique involves the use of an apparatus in which the digestive processes of the animal are conducted and can be studied in vitro. By this means, various animal feeds are introduced into or withdrawn from the laboratory unit under carefully controlled conditions and the changes taking place studied critically and progressively during the consumption of the feed by the microorganisms.

The results obtained by use of the aforesaid transplanted rumen technique have been confirmed by actual feeding of animals. This technique has established, for example, that protein addition to the feed substantially increases the consumption of cellulose by the animals and that also an increase in usable nitrogen to supplement protein feeds likewise improves ability to consume cellulose. The aforesaid and other results were determined first by laboratory experiments working in vitro and were later established in vivo by direct feeding of animals.

The feed supplements of this invention are the organic acid amides which not only supply a source of biologically available nitrogen as a nutrient for rumen microflora but also give a limited evolution of ammonia during ingestion. The low ammonia evolution determined in vitro is an assurance that these feed supplements can be used with wide latitudes in the composition of feeds. The carbon chain of these materials can be utilized by the microflora for their metabolic needs. In the examples which follow and which illustrate preferred embodiments of the invention, parts are by weight unless otherwise indicated.

This procedure was used in carrying out the processes of the examples. Into an Erlenmeyer flask of approximately 1 liter capacity was introduced 9 grams of cellulose, 0.6 gram of molasses ash, cc. of a nutrient salt solution, 1 gram of glucose and 450 cc. of a rumen inoculum, the mixture being diluted to 900 cc. with distilled water.

Sodium chloride, gm 7.50 Magnesium sulfate, gm; 2.25 Calcium chloride, gm 0.75 Ferrous sulfate, gm 0.15

Rumen inoculum was obtained from the first stomach of a fistulated steer which had been fed with a diet consisting of grain concentrate, alfalfa hay and salt mixture. The extracted rumen content was filtered through four thicknesses of cheese cloth or muslin of comparable porosity to remove solid matter, the liquid filtrate being" employed as the rumen inoculum. During the removal and filtration of the rumen contents care was exercised to exclude air. 1

The Erlenmeyer flask was charged with 450 cc. of this rumen liquid to which was added 450 cc.- of a distilled water suspension or solution of the aforementioned basal medium plus the nutrient to be evaluated. This flask was placed in a constant temperature water bath maintained at approximately 39 C. and carbon dioxide pumped through the charge in the flask to maintain anaerobic conditions.

At the end of the first 24 hours of bacterial digestion a sample was taken for analysis to determine ammonia, cellulose digestion and bacterial growth. After 24 hours digestion new nutrients in water were inoculated with half (450 cc.) of the previous days fermentation and diluted to 900 ml. with water. The analyses and subculturing into new nutrients was repeated over a 4 day period.

In the examples which follow comparisons are made between organic acid amides and urea. These comparisons show the cellulose digestion as well as the ammonia evolution of the compositions examined compared with a blank composition containing only cellulose, molasses ash, nutrient salt solution and rumen inoculum.

Example 1 FLASK CONTENTS Cellulose, g 9 Molasses ash, g. Nutrient salt soln., ml. 10% dextrose, ml.-.

HEIGHT OF NHa (Mg.)

o coco Example 2 FLASK CONTENTS Cellulose, g 9 9 9 10% dextrose s0ln., ml. 10 10 10 Molasses ash, g 0.6 0. 6 0.6 70 70 70 0.4

dilute to 900 m1.

HEIGHT OF NHa OELLULOSE DIGESTION (PERCENT) Example 3 FLASK CONTENTS Cellulose, g 9 9 9 Molasses ash, g 0.6 0.6 0.6 Nutrient salt s0ln., ml 70 70 70 10% dextrose s0ln., mL. 10 10 10 Urea, g 0.4 l- Glycinamide, HCL, g. 0.738 Rumen inoeulum, ml. .4--. 450 450 4 ,0 dilute to 900 m1.

HEIGHT 0F NH: (Mg.)

CELLULOSE DIGESTION (PERCENT) Example 4 FLASK CONTENTS Cellulose, Q 9 9 9 Molasses ash, g 0. 6 0.6 0.6 Nutrient salt soln., ml. 70 70 70 10% Dextrose, ml 10 10 10 Stare g 1 1 1 Ure 0.4 Formamide, g 0 599 Rumen inoculurn, ml 450 450 450 E 0. dilute 1:0900 ml.

HEIGHT 01* NH: (Mg.)

At end of:

1st; day 53 67 66 2nd day 18 87 44 3rd day 12 78 79 4th day 13 56 56 Av r- 24 72 61 The examples illustrate by way of comparison the ammonia evolved and the cellulose digested over a four day period. As the inoculum varies from day to day the comparison in each separate example, which generally was made on different days, necessitated the comparison of the nitrogen supplements with an inoculum taken for that day. It will be noted also that the comparisons are also made with an inoculum containing .4 gram of urea.

In Example 1 a propionamide inoculum is compared with inoculum with and without urea. It will be noted that an average of 139 mg. of ammonia was evolved from the urea while from propionamide an average of only 18 mg. was evolved. The cellulose digestion for urea averaged 59% while that of the propionamide averaged 56%.

In Example 2 an acetamide containing inoculum is similarly compared with an inoculum that contains urea and one that does not. Here the urea treated inoculum gave an average ammonia evolution of 135 mg. while that treated with acetamide gave only 77 mg. The urea treated inoculum averaged 88% cellulose digestion while that of acetamide 48%.

In Example 3 a glycinamide HCl treated inoculum is compared with an inoculum in the presence and absence of urea. In accord with this example the urea gave an average of 181 mg. of ammonia evolved while the glycinamide gave no ammonia evolution. The cellulose digestion for the urea treated inoculum is 85% and for the glycinamide treated inoculum 57%.

In Example 4 a formamide containing inoculum is compared with an inoculum that contains urea and one that does not. Here the ammonia level from the control and from the inoculum containing urea and formamide average respectively 12 m., 121 mg., and 55 mg., while the cellulose digestion average respectively 24%, 72%, and 61%.

The organic acid amides that may be used in accord with the invention, as nitrogen supplements for feeding rumen microflora and through them aid in cellulose digestion for Ruminantia, include the patentable non-toxic organic amides such, for example, as formamide, acetamide, propionamide, butyramide, stearamide, acrylamide, maleamide, oleamide, oxamide, glutaramide, succinimide, adipamide, maliamide, glycolamide, tartaramide, citramide, gluconamide, lactamide, sucramide, and glycinamide.

In the formulation of feeds for cattle and sheep, the organic acid amides can be used to supply from 5 to 90% of the nitrogen content of the protein requirements for the animal. This is a general rule that can be followed for complete rations for the production of protein concentrate mixtures or for the preparation of pelleted products. In the preparation of such formulated feeds, it is desirable that the synthetic nitrogen constituents thereof be preferably dry mixed with the other feed ingredients so that they can be evenly distributed throughout the feed. Any suitable mixture may be employed as, for example, a 14% protein feed can be made into an 18% feed composition by the addition of slightly more than 41 pounds of formamide per ton of feed. The usual type of feed composition is employed to provide the normal amount of carbohydrate content which may be supplied by Wheat, oats, barley, corn, hominy, molasses, and the like. These feeds may also contain oil meals and similar meals to supply natural proteins. For example, a typical 20% dairy ration for cattle may contain 300 lbs. of wheat bran, 310 lbs. of hominy feed, 200 lbs. of crimped oats, 300 lbs. of distillers dried grains, 175 lbs. of corn gluten feed, 400 lbs. of cocoanut oil meal, lbs. of soybean oil meal, 150 lbs. of molasses, 30 lbs. of formamide, 70 lbs. of brewers dried grains and 50 lbs. of minerals and salt.

Cattle are usually supplied with a diet of grass, hay and the like, augmented by a food concentrate such as that described in the last paragraph. These feed concentrates may contain from 10 to 50% protein, a small amount of minerals and the remainder mainly carbohydrate. The

minerals may be added directly to the feed concentrate. When preparing the concentrate in this manner such minerals as, e. g., defiuorinated phosphate rock, salt and the like may be used. Contrariwise, the minerals may be indirectly added with the carbohydrate and/or protein as a normal part of those ingredients. In preparing such feed concentrates it has been found that the amides hereinbefore described may constitute from 5 to of the nitrogen content of the feed, the remainder of the nitrogen being provided by protein.

The invention is not limited to the specific compositions described, nor to their specific proportions for the rate of utilization and its eificiency, as will be appreciated by those skilled in the art, is determined in large measure by the availability of the nitrogen to the rumen microorganisms and as a consequence, the percentage ingestion of a palatable amide of an organic acid that can be used varies over a wide range in accordnce with the nitrogen content of that feed supplement.

We claim:

1. A rumen microflora feed composition containing carbohydrates, protein and as a supplementary source of nitrogen an amide of an organic acid, the carbonyl group of said acid being attached to no more than one amido group.

2. A ruminant feed composition suitable for in vivo and in vitro feeding of rumen microflora containing an amide of an aliphatic organic acid, the carbonyl group of said acid being attached to no more than one amido group.

3. A ruminant feed composition containing carbohydrates, protein and as a supplementary source of nitrogen an amide of an aliphatic organic acid, the carbonyl group of said acid being attached to no more than one amido group.

4. A ruminant feed composition containing carbohydrates, minerals, protein and as a supplementary source of nitrogen an amide of an aliphatic organic acid, the carbonyl group of said acid being attached to no more than one amido group containing biologically available nitrogen which supplies from 5 to 90% of the nitrogen content of the feed.

5. A ruminant feed composition containing from 10 to 50% protein, minerals and the remainder mainly carbohydrate products, an amide of an aliphatic organic acid the carbonyl group of said acid being attached to no more than one amido group containing biologically available nitrogen supplying from 5 to 90% of the nitrogen content of the feed.

6. A ruminant feed composition suitable for the in vivo and in vitro feeding of rumen microflora containing carbohydrates, protein and, as a supplementary source of nitrogen, propionamide.

7. A ruminant feed composition suitable for the in vivo and in vitro feeding of rumen microflora containing carbohydrates, protein and, as a supplementary source of nitrogen, acetamide.

8. A ruminant feed composition suitable for the in vivo and in vitro feeding of rumen microflora containing carbohydrates, protein and, as a supplementary source of nitrogen, glycinamide.

9. A ruminant feed composition suitable for the in vivo and in vitro feeding of rumen microflora containing carbohydrates, protein and, as a supplementary source of nitrogen, forrnamide.

10. A ruminant feed composition suitable for the in vivo and in vitro feeding of rumen microflora containing carbohydrates, protein and, as a supplementary source of nitrogen, lactamide.

References Cited in the file of this patent UNITED STATES PATENTS 2,560,830 Turner et a1 July 17, 1951 

1. A RUMEN MICROFLORA FEED COMPOSITON CONTAINING CARBOHYDRATES, PROTEIN AND AS A SUPPLEMENTARY SOURCE OF NITROGEN AN AMIDE OF AN ORGANIC ACID, THE CARBONYL GROUP OF SAID ACID BEING ATTACHED TO NO MORE THAN ONE AMIDO GROUP. 